The goal of this research is to characterize the role of Treponema denticola (Td) surface proteins in interaction of this human oral spirochete with host tissue, thereby gaining insight into mechanisms by which these organisms contribute to initiation and progression of periodontal disease. We focus on analysis of Td protein complexes that directly affect cells isolated from tissue comprising the periodontal ligament (PDL) that comprises the junction between the tooth and the alveolar bone of the tooth socket: specifically the PrtP lipoprotein protease complex (dentilisin) and the oligomeric Msp protein. Our overall hypothesis is that dentilisin and Msp are major contributors to Td cytopathic behavior in periodontal disease. To characterize their specific roles in microbe-host interactions, our approach is to utilize purified native and recombinant proteins as well as isogenic Td strains carrying defined mutations in individual components of these outer membrane complexes. We will first extend our ongoing studies characterizing dentilisin assembly in the Td outer membrane. Then, to further studies of host cell responses to Td challenge, we will (A) determine the mechanism of dentilisin-induced activation of pro-MMP-2 and (B) characterize MMP-2-dependent fibronectin fragmentation resulting from Td challenge of PDL cells. This project will test the hypothesis that dentilisin is an effector protein whose activity results in magnified downstream signaling effects that are likely to be significant in vivo. Our research team is uniquely positioned to conduct these studies, with combined expertise in spirochete molecular biology, extracellular matrix biology and cytopathology of inflammatory diseases. Completion of this project will contribute to both basic knowledge of spirochete molecular biology and to understanding of microbe-host interactions in chronic infections such as periodontal diseases.